Starch branching enzyme II of potato

ABSTRACT

The present invention relates to an amino acid sequence of second starch branching enzyme (SBE II) of potato and a fragment thereof as well as to the corresponding isolated DNA sequences. Furthermore, the invention relates to vectors comprising such an isolated DNA sequence, to processes for production of transgenic potatoes, and to the use of said potatoes for the production of starch. The starch obtained will show a changed pattern of branching of amylopectin as well as a changed amylose/amylopectin ratio.

This is a continuation of International Application No. PCT/SE96/01558,filed Nov. 28, 1996 that designates the United States.

FIELD OF THE INVENTION

The present invention relates to a novel starch branching enzyme ofpotato. More specifically, the present invention relates to an aminoacid sequence of a second starch branching enzyme (SBE II) of potato anda fragment thereof as well as their corresponding DNA sequences.Furthermore, the invention relates to vectors comprising such DNAsequences, to processes for production of transgenic potatoes, and tothe use of said potatoes for the production of starch.

Starch is a complex mixture of different molecule forms differing indegree of polymerization and branching of the glucose chains. Starchconsists of amylose and amylopectin, whereby the amylose consists of anessentially linear α-1,4-glucan and amylopectin consists ofα-1,4-glucans connected to each other via α-1,6-linkages and, thus,forming a branched polyglucan. Thus, starch is not a uniform rawmaterial.

Starch is synthesized via at least three enzymatic reactions in whichADP glucose phosphorylase (EC 2.7.7.27), starch synthase (EC 2.4.1.21)and starch branching enzyme (EC 2.4.1.18) are involved. Starch branchingenzyme (SBE, also called Q-enzyme) is believed to have two differentenzymatic activities. It catalyzes both the hydrolysis ofα-1,4-glucosidic bonds and the formation of α-1,6-glucosidic bondsduring synthesis of the branched component in starch, i.e. amylopectin.

Plant starch is a valuable source of renewable raw material used in, forexample, the chemical industry (Visser and Jacobsen, 1993). However, thequality of the starch has to meet the demands of the processing industrywherein uniformity of structure is an important criterion. Forindustrial application there is a need of plants only containing amylosestarch and plants only containing amylopectin starch, respectively.

Processes for altering the amylose/amylopectin ratio in starch havealready been proposed. For example, in WO95/04826 there is described DNAsequences encoding debranching enzymes with the ability to reduce orincrease the degree of branching of amylopectin in transgenic plants,e.g. potatoes.

In WO92/14827 plasmids are described having DNA sequences that afterinsertion into the genome of the plants cause changes in thecarbohydrate concentration and the carbohydrate composition inregenerated plants. These changes can be obtained from a sequence of abranching enzyme that is located on these plasmids. This branchingenzyme is proposed to alter the amylose/amylopectin ratio in starch ofthe plants, especially in commercially used plants.

WO92/14827 describes the only hitherto known starch branching enzyme inpotato and within the art it is not known whether other starch branchingenzymes are involved in the synthesis of branched starch of potato.

In Mol Gen Genet (1991) 225:289-296, Visser et al., there is describedinhibition of the expression of the gene for granule-bound starchsynthase in potato by antisense constructs. Inhibition of the enzyme inpotato tuber starch was up to 100% in which case amylose-free starch wasprovided.

However, the prior known methods for inhibiting amylopectin have notbeen that successful and, therefore, alternative methods for inhibitingamylopectin are still highly desirable (M{umlaut over (u)}ller-Röber andKoβmann, 1994; Martin and Smith, 1995).

SUMMARY OF THE INVENTION

The object of the present invention is to enable altering the degree ofamylopectin branching and the amylopectin/amylose ratio in potatostarch.

According to the present invention this object is achieved by providinga novel isolated DNA sequence encoding a second starch branching enzyme,SBE II, and fragments thereof, which after insertion into the genome ofthe plants cause changes in said branching degree and ratio inregenerated plants.

Within the scope of the present invention there is also included theamino acid sequence of SBE II and fragments thereof.

Also variants of the above DNA sequence resulting from the degeneracy ofthe genetic code are encompassed.

The novel DNA sequence encoding SBEII, comprising 3074 nucleotides, aswell as the corresponding amino acid sequence comprising 878 aminoacids, are shown in SEQ ID No. 1. One 1393 nucleotides long fragment ofthe above DNA sequence, corresponding to nucleotides 1007 to 2399 of theDNA sequence in SEQ ID No. 1, as well as the corresponding amino acidsequence comprising 464 amino acids, are shown in SEQ ID No. 2.

Furthermore, there are provided vectors comprising said isolatedDNA-sequences and regulatory elements active in potato. The DNAsequences may be inserted in the sense or antisense (reversed)orientation in the vectors in relation to a promoter immediatelyupstream from the DNA sequence.

Also there is provided a process for the production of transgenicpotatoes with a reduced degree of branching of amylopectin starch,comprising the following steps:

a) transfer and incorporation of a vector according to the inventioninto the genome of a potato cell, and

b) regeneration of intact, whole plants from the transformed cells.

Finally, the invention provides the use of said transgenic potatoes forthe production of starch.

The invention will be described in more detail below in association withan experimental part and the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows SDS polyacrylamide electrophoresis of proteins extractedfrom starch of normal potato (lane A) and transgenic potato (lane B).Excised protein bands are marked with arrows. Lane M: Molecular weightmarker proteins (kDa).

FIG. 2 shows 4 peptide sequences derived from digested proteins frompotato tuber starch.

DETAILED DESCRIPTION OF THE INVENTION

Isolation of starch from potato tubers

Potato plants (Solanum tuberosum) were grown in the field. Peeled tubersfrom either cv. Early Puritan or from a transgenic potato lineessentially lacking granule-bound starch synthase I (Svalöf Weibull AB,international application number PCT/SE91/00892), were homogenized at 4°C. in a fruit juicer. To the “juice fraction”, which contained a largefraction of the starch, was immediately added Tris-HCl, pH 7.5, to 50mM, Na-dithionite to 30 mM and ethylenedinitrilotetraacetic acid (EDTA)to 10 mM. The starch granules were allowed to sediment for 30 min andwashed 4× with 10 bed volumes of washing buffer (50 mM Tris-HCl, pH 7.5,10 mM EDTA). The starch, which was left on the bench at +4° C. for 30min to sediment between every wash, was finally washed with 3×3 bedvolumes of acetone, air dried over night, and stored at −20° C.

Extraction of proteins from tuber starch

Stored starch (20 g) was continuously mixed with 200 ml extractionbuffer (50 mM Tris-HCl, pH 7.5, 2% (w/v) sodium dodecyl sulfate (SDS), 5mM EDTA) by aspiration with a pipette at 85° C. until the starch wasgelatinized. The samples were then frozen at −70° C. for 1 hour. Afterthawing at 50° C., the samples were centrifuged for 20 min at 12,000×gat 10° C. The supernatants were collected and re-centrifuged at 3,000×gfor 15 min. The final supernatants were filtered through 0.45μ filtersand 2.25 volumes of ice-cold acetone were added. After 30 min incubationat 4° C., the protein precipitates were collected by centrifugation(3,000×g for 30 min at 4° C.), and dissolved in 50 mM Tris-HCl, pH 7.5.An aliquot of each preparation was analyzed by SDS poly-acrylamide gelelectrophoresis according to Laemmli (1970) (FIG. 1). The proteins inthe remaining portions of the preparations were concentrated byprecipitation with trichloroacetic acid (10%) and the proteins wereseparated on an 8% SDS polyacrylamide gel Laemmli, (1970). The proteinsin the gel were stained with Coomassie Brilliant Blue R-250 (0.2% in 20%methanol, 0.5% acetic acid, 79.5% H₂O).

In gel digestion and sequencing of peptides

The stained bands marked with arrows in FIG. 1 corresponding to anapparent molecular weight of about 100 kDa were excised and washed twicewith 0.2M NH₄HCO₃ in 50% acetonitrile under continuous stirring at 35°C. for 20 min. After each washing, the liquid was removed and the gelpieces were allowed to dry by evaporation in a fume hood. The completelydried gel pieces were then separately placed on parafilm and 2 μl of0.2M NH₄CO₃, 0.02% Tween-20 were added. Modified trypsin (Promega,Madison, Wis., USA) (0.25 μg in 2 μl) was sucked into the gel pieceswhereafter 0.2M NH₄CO₃ was added in 5 μl portions until they had resumedtheir original sizes. The gel slices were further divided into threepieces and transferred to an Eppendorf tube. 0.2M NH₄CO₃ (200 μl) wasadded and the proteins contained in the gel pieces were digested overnight at 37° C. (Rosenfeld et al. 1992). After completed digestion,trifluoroacetic acid was added to 1% and the supernatants removed andsaved. The gel pieces were further extracted twice with 60%acetonitrile, 0.1% trifluoroacetic acid (200 μl) under continuousshaking at 37° C. for 20 min. The two supernatants from theseextractions were combined with the first supernatant. The gel pieceswere finally washed with 60% acetonitrile, 0.1% trifluoroacetic acid,0.02% Tween-20 (200 μl). Also these supernatants were combined with theother supernatants and the volume was reduced to 50 μl by evaporation.The extracted peptides were separated on a SMART® chromatography system(Pharmacia, Uppsala, Sweden) equipped with a μRPC C2/C18 SC2.1/10column. Peptides were eluted with a gradient of 0-60% acetonitrile inwater/0.1% trifluoroacetic acid over 60 min with a flow rate of 100μl/min. Peptides were sequenced either on an Applied Biosystems 470A gasphase sequenator with an on line PTH-amino acid analyzer (120A) or on amodel 476A according to the instructions of the manufacturer (AppliedBiosystems, Foster City, Calif., USA).

Four of the peptides sequenced gave easily interpretable sequences (FIG.2). A data base search revealed that these four peptides displayedsimilarity to starch branching enzymes and interestingly, the peptideswere more related to starch branching enzyme II from other plant speciesthan to starch branching enzyme I from potato.

Construction of oligonucleotides encoding peptides 1 and 2.

Degenerated oligonucleotides encoding peptide 1 and peptide 2 weresynthesized as forward and reverse primers, respectively:

Oligonucleotide 1: 5′-gtaaaacgacggccagt-TTYGGNGTNTGGGARATHTT-3′(Residues 2 to 8 of peptide 1)

Oligonucleotide 2: 5′-aattaaccctcactaaaggg-CKRTCRAAYTCYTGIARNCC-3′(Residues 2 to 8 of peptide 2, reversed strand) wherein

H is A, C or T, I is inosine; K is G or T; N is A, C, G or T; R is A orG; Y is C or T; bases in lower case were added as tag sequences.

Purification of mRNA from potato tuber, synthesis of cDNA and PCRamplification of a cDNA fragment corresponding to potato starchbranching enzyme II.

Total RNA from mature potato tubers (S. tuberosum cv. Amanda) wasisolated as described (Logemann et al. 1987). First strand cDNA wassynthesized using 2 μg of total RNA and 60 pmol of oligo-dT₃₀ asdownstream primer. The primer was annealed to the polyA of the mRNA at60° C. for 5 min. The extension of the cDNA was performed according tothe technical manual of the manufacturer using the Riboclone® cDNASynthesis System M-MLV (H−) (Promega).

cDNA encoding the novel starch branching enzyme II according to theinvention was amplified in a Perkin-Elmer GeneAmp® 9600 PCR thermocycler(Perkin-Elmer Cetus Instruments, Conn., USA) using the two degenerateprimers designed from the peptides 1 and 2 (see above) under thefollowing conditions: 1 mM dNTP, 1 μM of each primer and an alicot ofthe cDNA described above in a total reaction volume of 20 μl with 1×AmpliTaq® buffer and 0.8 U AmpliTaq® (Perkin-Elmer Cetus). The cyclingconditions were: 96° C. for 1′, 80° C. while the enzyme was added as ahotstart (approximately 15′), an unintended drop to 25° C., five cyclesof 94° C. for 20″, 45° C. for 1′, ramp to 72° C. for 1′ and 72° C. for2′, and 30 cycles of 94° C. for 5″, 45° C. for 30″, and 72° C. for(2′+2″ per cycle) and completed with 72° C. for 10′ prior to chilling to4° C.

A sample of this reaction (0.1 μl) was reamplified using the cyclingconditions: 96° C. for 1′, 80° C. while the enzyme was added as ahotstart (approximately 5′), five cycles of 94° C. for 20″, 45° C. for1′, and 72° C. for 2′, and 25 cycles of 94° C. for 5″, 45° C. for 30″,and 72° C. for (2′+2″ per cycle) and completed with 72° C. for 10′ priorto chilling to 4° C. After completion of the PCR amplification, thereaction was loaded on a 1.5% Seakem® agarose gel (FMC Bioproducts,Rockland, Me., USA). After electrophoresis and staining with ethidiumbromide a major band with an apparent size of 1500 bp was excised andthe fragment was eluted by shaking in water (200 μl) for 1 h. Thisfragment was used as template in sequencing reactions afterreamplification using primers corresponding to the tag sequences (inoligonucleotides 1 and 2), purification by agarose gel electrophoresisas above and extraction from the gel using the Qiaex® gel extraction kitaccording to the manufacturer's instructions (DIAGEN GmbH, Hilden,Germany). The sequencing reactions were done using the DyeDeoxy®Terminator Cycle Sequencing kits (Perkin-Elmer Cetus Instruments) usingtag sequences and internal primers. The sequencing reaction wereanalyzed on an Applied Biosystems 373A DNA sequencer according to themanufacturer's protocol. The sequence was edited and comprised 1393 bp.

To complete the determination of the sequence of starch branching enzymeII, the 5′ and 3′ ends of the full length cDNA were amplified from thesame total RNA as above using rapid amplification of cDNA ends, RACEmethodology with specific primers from the 1393 bp sequence. In the 3′end amplification, an oligo T₂₉G primer was used against the poly A tailand in the 5′ end, the 5′/3′ RACE kit from Boehringer Mannheim (Cat. No.1734792) was used. The fragments from these amplifications weresequenced in the same way as above using internal and end primers. Thesequences from the two ends were aligned together with the 1393 basepairs to give a composite full length cDNA sequence. Primers weredesigned from this sequence to amplify the whole coding region in onepart. Partial sequencing of the amplified coding cDNA confirmed thepresence of a cDNA corresponding to the composite sequence. The fulllength cDNA is 3074 bp and the translated sequence comprises 878 aminoacids. The mature protein comprises 830 amino acids.

Comparisons of the consensus sequence with the EMBL and GenBankdatabases showed 68% identity to potato starch branching enzyme I andabout 80% identity to starch branching enzyme II from other plantspecies. The present inventors therefore denote the enzyme encoded bythe new branching enzyme sequence potato starch branching enzyme II.

Transformation of potato plants

The isolated full length cDNA of potato starch branching enzyme II andother functionally active fragments in the range of 50-3 074 bp arecloned in reverse orientation behind promoters active in potato tubers.By the term “functionally active” is meant fragments that will affectthe amylose/amylopectin ratio in potato starch. The DNA and amino acidsequence of SBE II according to the invention as well as one fragment ofthe DNA and corresponding amino acid sequence are shown in SEQ ID No. 1and 2, respectively.

The promoters are selected from, for example, the patatin promoter, thepromoter from the potato granule-bound starch synthase I gene orpromoters isolated from potato starch branching enzymes I and II genes.

The constructs are cloned by techniques known in the art either in abinary Ti-plasmid vector suitable for transformation of potato mediatedby Agrobacterium tumefaciens, or in a vector suitable for directtransformation using ballistic techniques or electroporation. It isrealized that the sense (see below) and antisense constructs mustcontain all necessary regulatory elements.

Transgenic potato plants transcribe the inverse starch branching enzymeII construct specifically in tubers, leading to antisense inhibition ofthe enzyme. A reduction and changed pattern of the branching ofamylopectin as well as a changed amylose/amylopectin ratio thereby occurin tuber starch.

The antisense construct for potato starch branching enzyme II is alsoused in combination with antisense constructs for potato starchbranching enzyme I, for potato granule-bound starch synthase II, forpotato soluble starch synthases II and III, for potato starchdisproportionating enzyme (D-enzyme) or for potato starch debranchingenzyme to transform potato to change the degree of branching ofamylopectin and the amylose/amylopectin ratio. This gives new andvaluable raw material to the starch processing industry.

The full-length cDNA sequence encoding the enzyme is, in differentconstructs, cloned in sense orientation behind one or more of thepromoters mentioned above, and the constructs are transferred intosuitable transformation of potato. Regenerated transformed potato plantswill produce an excess of starch branching enzyme II in the tubersleading to an increased degree and changed pattern of branching ofamylopectin or to inhibition of transcription of endogenous starchbranching enzyme II transcription due to co-suppression, resulting in adecreased branching or amylopectin.

References

M{umlaut over (u)}ller-Röber, B., Koβmann, J., (1994) Approaches toinfluence starch quantity and starch quality in transgenic plants. PlantCell Environm. 17, 601-613.

Martin, C., Smith, A. (1995) Starch Biosynthesis. Plant Cell 7, 971-985.

Laemmli, U. K. (1979) Cleavage of structural proteins during assembly ofthe head of bacteriophage T4. Nature 227, 680-685.

Logemann, J., Schell, J. and Willmitzer, L. (1987) Improved method forthe isolation of RNA from plant tissues. Anal. Biochem. 163, 16-20.

Rosenfeld, J., Capdeville, J, Guillemot, J. C., Ferrara, P. (1992)In-gel digestion of proteins for internal sequence analysis after one-or two-dimensional gel electrophoresis. Anal. Biochem. 203, 173-179.

Visser, R. G. F., Jacobsen, E. (1993) Towards modifying plants foraltered starch content and composition. TibTech 11, 63-68.

4 1 3074 DNA Unknown Description of Unknown OrganismbeII gene (branchingenzyme II) from Solanum tuberosum (potato) 1 aaacctcctc cactcagtctttgtttctct ctctcttcac gcttctcttg gcgccttgaa 60 ctcagcaatt tgacactcagttagttacac tnccatcact tatcagatct ctattttttc 120 tcttaattcc aaccaaggaatgaataaaaa gatagatttg taaaaaccct aaggagagaa 180 gaagaaag atg gtg tat acactc tct gga gtt cgt ttt cct act gtt cca 230 Met Val Tyr Thr Leu Ser GlyVal Arg Phe Pro Thr Val Pro -45 -40 -35 tca gtg tac aaa tct aat gga ttcagc agt aat ggt gat cgg agg aat 278 Ser Val Tyr Lys Ser Asn Gly Phe SerSer Asn Gly Asp Arg Arg Asn -30 -25 -20 gct aat ntt tct gta ttc ttg aaaaag cac tct ctt tca cgg aag atc 326 Ala Asn Xaa Ser Val Phe Leu Lys LysHis Ser Leu Ser Arg Lys Ile -15 -10 -5 ttg gct gaa aag tct tct tac aattcc gaa tcc cga cct tct aca gtt 374 Leu Ala Glu Lys Ser Ser Tyr Asn SerGlu Ser Arg Pro Ser Thr Val -1 1 5 10 gca gca tcg ggg aaa gtc ctt gtgcct gga acc cag agt gat agc tcc 422 Ala Ala Ser Gly Lys Val Leu Val ProGly Thr Gln Ser Asp Ser Ser 15 20 25 30 tca tcc tca aca gac caa ttt gagttc act gag aca tct cca gaa aat 470 Ser Ser Ser Thr Asp Gln Phe Glu PheThr Glu Thr Ser Pro Glu Asn 35 40 45 tcc cca gca tca act gat gta gat agttca aca atg gaa cac gct agg 518 Ser Pro Ala Ser Thr Asp Val Asp Ser SerThr Met Glu His Ala Arg 50 55 60 cag att aaa act gag aac gat gac gtt gagccg tca agt gat ctt aca 566 Gln Ile Lys Thr Glu Asn Asp Asp Val Glu ProSer Ser Asp Leu Thr 65 70 75 gga agt gtt gaa gag ctg gat ttt gct tca tcacta caa cta caa gaa 614 Gly Ser Val Glu Glu Leu Asp Phe Ala Ser Ser LeuGln Leu Gln Glu 80 85 90 ggt ggt aaa ctg gag gag tct aaa aca tta aat acttct gaa gag aca 662 Gly Gly Lys Leu Glu Glu Ser Lys Thr Leu Asn Thr SerGlu Glu Thr 95 100 105 110 att att gat gaa tct gat agg atc aga gag aggggc atc cct cca cct 710 Ile Ile Asp Glu Ser Asp Arg Ile Arg Glu Arg GlyIle Pro Pro Pro 115 120 125 gga ctt ggt cag aag att tat gaa ata gac cccctt ttg aca aac tat 758 Gly Leu Gly Gln Lys Ile Tyr Glu Ile Asp Pro LeuLeu Thr Asn Tyr 130 135 140 cgt caa cac ctt gat tac agg tat tca cag tacaag aaa ctg agg gag 806 Arg Gln His Leu Asp Tyr Arg Tyr Ser Gln Tyr LysLys Leu Arg Glu 145 150 155 gca att gac aag tat gag ggt ggt ttg gaa gctttt tct cgt ggt tat 854 Ala Ile Asp Lys Tyr Glu Gly Gly Leu Glu Ala PheSer Arg Gly Tyr 160 165 170 gaa aaa atg ggt ttc act cgt agt gct aca ggtatc act tac cgt gag 902 Glu Lys Met Gly Phe Thr Arg Ser Ala Thr Gly IleThr Tyr Arg Glu 175 180 185 190 tgg gct cct ggt gcc cag tca gct gcc ctcatt gga gat ttc aac aat 950 Trp Ala Pro Gly Ala Gln Ser Ala Ala Leu IleGly Asp Phe Asn Asn 195 200 205 tgg gac gca aat gct gac att atg act cggaat gaa ttt ggt gtc tgg 998 Trp Asp Ala Asn Ala Asp Ile Met Thr Arg AsnGlu Phe Gly Val Trp 210 215 220 gag att ttt ctg cca aat aat gtg gat ggttct cct gca att cct cat 1046 Glu Ile Phe Leu Pro Asn Asn Val Asp Gly SerPro Ala Ile Pro His 225 230 235 ggg tcc aga gtg aag ata cgt atg gac actcca tca ggt gtt aag gat 1094 Gly Ser Arg Val Lys Ile Arg Met Asp Thr ProSer Gly Val Lys Asp 240 245 250 tcc att cct gct tgg atc aac tac tct ttacag ctt cct gat gaa att 1142 Ser Ile Pro Ala Trp Ile Asn Tyr Ser Leu GlnLeu Pro Asp Glu Ile 255 260 265 270 cca tat aat gga ata tat tat gat ccaccc gaa gag gag agg tat atc 1190 Pro Tyr Asn Gly Ile Tyr Tyr Asp Pro ProGlu Glu Glu Arg Tyr Ile 275 280 285 ttc caa cac cca cgg cca aag aaa ccaaag tcg ctg aga ata tat gaa 1238 Phe Gln His Pro Arg Pro Lys Lys Pro LysSer Leu Arg Ile Tyr Glu 290 295 300 tct cat att gga atg agt agt ccg gagcct aaa att aac tca tac gtg 1286 Ser His Ile Gly Met Ser Ser Pro Glu ProLys Ile Asn Ser Tyr Val 305 310 315 aat ttt aga gat gaa gtt ctt cct cgcata aaa aag ctt ggg tac aat 1334 Asn Phe Arg Asp Glu Val Leu Pro Arg IleLys Lys Leu Gly Tyr Asn 320 325 330 gcg gtg caa att atg gct att caa gagcat tct tat tat gct agt ttt 1382 Ala Val Gln Ile Met Ala Ile Gln Glu HisSer Tyr Tyr Ala Ser Phe 335 340 345 350 ggt tat cat gtc aca aat ttt ttngca cca agc agc cgt ttt gga acn 1430 Gly Tyr His Val Thr Asn Phe Xaa AlaPro Ser Ser Arg Phe Gly Xaa 355 360 365 ccc gac gac ctt aag tct ttg attgat aaa gct cat gag cta gga att 1478 Pro Asp Asp Leu Lys Ser Leu Ile AspLys Ala His Glu Leu Gly Ile 370 375 380 gtt gtt ctc atg gac att gtt cacagc cat gca tca aat aat act tta 1526 Val Val Leu Met Asp Ile Val His SerHis Ala Ser Asn Asn Thr Leu 385 390 395 gat gga ctg aac atg ttt gac ggcaca gat agt tgt tac ttt cac tct 1574 Asp Gly Leu Asn Met Phe Asp Gly ThrAsp Ser Cys Tyr Phe His Ser 400 405 410 gga gct cgt ggt tat cat tgg atgtgg gat tcc cgc ctc ttt aac tat 1622 Gly Ala Arg Gly Tyr His Trp Met TrpAsp Ser Arg Leu Phe Asn Tyr 415 420 425 430 gga aac tgg gag gta ctt aggtat ctt ctc tca aat gcg aga tgg tgg 1670 Gly Asn Trp Glu Val Leu Arg TyrLeu Leu Ser Asn Ala Arg Trp Trp 435 440 445 ttg gat gag ttc aaa ttt gatgga ttt aga ttt gat ggt gtg aca tca 1718 Leu Asp Glu Phe Lys Phe Asp GlyPhe Arg Phe Asp Gly Val Thr Ser 450 455 460 atg atg tat act cac cac ggatta tcg gtg gga ttc act ggg aac tac 1766 Met Met Tyr Thr His His Gly LeuSer Val Gly Phe Thr Gly Asn Tyr 465 470 475 gag gaa tac ttt gga ctc gcaact gat gtg gat gct gtt gtg tat ctg 1814 Glu Glu Tyr Phe Gly Leu Ala ThrAsp Val Asp Ala Val Val Tyr Leu 480 485 490 atg ctg gtc aac gat ctt attcat ggg ctt ttc cca gat gca att acc 1862 Met Leu Val Asn Asp Leu Ile HisGly Leu Phe Pro Asp Ala Ile Thr 495 500 505 510 att ggt gaa gat gtt agcgga atg ccg aca ttt tnt att ccc gtt caa 1910 Ile Gly Glu Asp Val Ser GlyMet Pro Thr Phe Xaa Ile Pro Val Gln 515 520 525 gat ggg ggt gtt ggc tttgac tat cgg ctg cat atg gca att gct gat 1958 Asp Gly Gly Val Gly Phe AspTyr Arg Leu His Met Ala Ile Ala Asp 530 535 540 aaa tgg att gag ttg ctcaag aaa cgg gat gag gat tgg aga gtg ggt 2006 Lys Trp Ile Glu Leu Leu LysLys Arg Asp Glu Asp Trp Arg Val Gly 545 550 555 gat att gtt cat aca ctgaca aat aga aga tgg tcg gaa aag tgt gtt 2054 Asp Ile Val His Thr Leu ThrAsn Arg Arg Trp Ser Glu Lys Cys Val 560 565 570 tca tac gct gaa agt catgat caa gct cta gtc ggt gat aaa act ata 2102 Ser Tyr Ala Glu Ser His AspGln Ala Leu Val Gly Asp Lys Thr Ile 575 580 585 590 gca ttc tgg ctg atggag aag gat atg tat gat ttt atg gct ctg gat 2150 Ala Phe Trp Leu Met GluLys Asp Met Tyr Asp Phe Met Ala Leu Asp 595 600 605 aga ccn tca aca tcatta ata gat cgt ggg ata gca ttg cac aag atg 2198 Arg Xaa Ser Thr Ser LeuIle Asp Arg Gly Ile Ala Leu His Lys Met 610 615 620 att agg ctt gta actatg gga tta gga gga gaa ggg tac cta aat ttc 2246 Ile Arg Leu Val Thr MetGly Leu Gly Gly Glu Gly Tyr Leu Asn Phe 625 630 635 atg gga aat gaa ttcggc cac cct gag tgg att gat ttc cct agg gct 2294 Met Gly Asn Glu Phe GlyHis Pro Glu Trp Ile Asp Phe Pro Arg Ala 640 645 650 gaa caa cac ctc tctgat ggc tca gta att ccc gga aac caa ttc agt 2342 Glu Gln His Leu Ser AspGly Ser Val Ile Pro Gly Asn Gln Phe Ser 655 660 665 670 tat gat aaa tgcaga cgg aga ttt gac ctg gga gat gca gaa tat tta 2390 Tyr Asp Lys Cys ArgArg Arg Phe Asp Leu Gly Asp Ala Glu Tyr Leu 675 680 685 aga tac cgt gggttg caa gaa ttt gac cgg gct atg cag tat ctt gaa 2438 Arg Tyr Arg Gly LeuGln Glu Phe Asp Arg Ala Met Gln Tyr Leu Glu 690 695 700 gat aaa tat gagttt atg act tca gaa cac cag ttc ata tca cga aag 2486 Asp Lys Tyr Glu PheMet Thr Ser Glu His Gln Phe Ile Ser Arg Lys 705 710 715 gat gaa gga gatagg atg att gta ttt gaa aaa gga aac cta gtt ttt 2534 Asp Glu Gly Asp ArgMet Ile Val Phe Glu Lys Gly Asn Leu Val Phe 720 725 730 gtc ttt aat tttcac tgg aca aaa agc tat tca gac tat cgc ata ggc 2582 Val Phe Asn Phe HisTrp Thr Lys Ser Tyr Ser Asp Tyr Arg Ile Gly 735 740 745 750 tgc ctg aagcct gga aaa tac aag gtt gcc ttg gac tca gat gat cca 2630 Cys Leu Lys ProGly Lys Tyr Lys Val Ala Leu Asp Ser Asp Asp Pro 755 760 765 ctt ttt ggtggc ttc ggg aga att gat cat aat gcc gaa tat ttc acc 2678 Leu Phe Gly GlyPhe Gly Arg Ile Asp His Asn Ala Glu Tyr Phe Thr 770 775 780 ttt gaa ggatgg tat gat gat cgt cct cgt tca att atg gtg tat gca 2726 Phe Glu Gly TrpTyr Asp Asp Arg Pro Arg Ser Ile Met Val Tyr Ala 785 790 795 cct agt agaaca gca gtg gtc tat gca cta gta gac aaa gaa gaa gaa 2774 Pro Ser Arg ThrAla Val Val Tyr Ala Leu Val Asp Lys Glu Glu Glu 800 805 810 gaa gaa gaagaa gta gca gta gta gaa gaa gta gta gta gaa gaa gaa 2822 Glu Glu Glu GluVal Ala Val Val Glu Glu Val Val Val Glu Glu Glu 815 820 825 830 tgaacgaacttgt gatcgcgttg aaagatttga aggctacata gagcttcttg 2875 acgtatctggcaatattgca tcagtcttgg cggaatttca tgtgacaaaa ggtttgcaat 2935 tctttccactattagtagtg caacgatata cgcagagatg aagtgctgca caaacatatg 2995 taaaatcgatgaatttatgt cgaatgctgg gacgggcttc agcaggtttt gcttagtgag 3055 ttctgtaaattgtcatctc 3074 2 878 PRT Unknown Description of Unknown OrganismbeIIgene (branching enzyme II) from Solanum tuberosum (potato) 2 Met Val TyrThr Leu Ser Gly Val Arg Phe Pro Thr Val Pro Ser Val 1 5 10 15 Tyr LysSer Asn Gly Phe Ser Ser Asn Gly Asp Arg Arg Asn Ala Asn 20 25 30 Xaa SerVal Phe Leu Lys Lys His Ser Leu Ser Arg Lys Ile Leu Ala 35 40 45 Glu LysSer Ser Tyr Asn Ser Glu Ser Arg Pro Ser Thr Val Ala Ala 50 55 60 Ser GlyLys Val Leu Val Pro Gly Thr Gln Ser Asp Ser Ser Ser Ser 65 70 75 80 SerThr Asp Gln Phe Glu Phe Thr Glu Thr Ser Pro Glu Asn Ser Pro 85 90 95 AlaSer Thr Asp Val Asp Ser Ser Thr Met Glu His Ala Arg Gln Ile 100 105 110Lys Thr Glu Asn Asp Asp Val Glu Pro Ser Ser Asp Leu Thr Gly Ser 115 120125 Val Glu Glu Leu Asp Phe Ala Ser Ser Leu Gln Leu Gln Glu Gly Gly 130135 140 Lys Leu Glu Glu Ser Lys Thr Leu Asn Thr Ser Glu Glu Thr Ile Ile145 150 155 160 Asp Glu Ser Asp Arg Ile Arg Glu Arg Gly Ile Pro Pro ProGly Leu 165 170 175 Gly Gln Lys Ile Tyr Glu Ile Asp Pro Leu Leu Thr AsnTyr Arg Gln 180 185 190 His Leu Asp Tyr Arg Tyr Ser Gln Tyr Lys Lys LeuArg Glu Ala Ile 195 200 205 Asp Lys Tyr Glu Gly Gly Leu Glu Ala Phe SerArg Gly Tyr Glu Lys 210 215 220 Met Gly Phe Thr Arg Ser Ala Thr Gly IleThr Tyr Arg Glu Trp Ala 225 230 235 240 Pro Gly Ala Gln Ser Ala Ala LeuIle Gly Asp Phe Asn Asn Trp Asp 245 250 255 Ala Asn Ala Asp Ile Met ThrArg Asn Glu Phe Gly Val Trp Glu Ile 260 265 270 Phe Leu Pro Asn Asn ValAsp Gly Ser Pro Ala Ile Pro His Gly Ser 275 280 285 Arg Val Lys Ile ArgMet Asp Thr Pro Ser Gly Val Lys Asp Ser Ile 290 295 300 Pro Ala Trp IleAsn Tyr Ser Leu Gln Leu Pro Asp Glu Ile Pro Tyr 305 310 315 320 Asn GlyIle Tyr Tyr Asp Pro Pro Glu Glu Glu Arg Tyr Ile Phe Gln 325 330 335 HisPro Arg Pro Lys Lys Pro Lys Ser Leu Arg Ile Tyr Glu Ser His 340 345 350Ile Gly Met Ser Ser Pro Glu Pro Lys Ile Asn Ser Tyr Val Asn Phe 355 360365 Arg Asp Glu Val Leu Pro Arg Ile Lys Lys Leu Gly Tyr Asn Ala Val 370375 380 Gln Ile Met Ala Ile Gln Glu His Ser Tyr Tyr Ala Ser Phe Gly Tyr385 390 395 400 His Val Thr Asn Phe Xaa Ala Pro Ser Ser Arg Phe Gly XaaPro Asp 405 410 415 Asp Leu Lys Ser Leu Ile Asp Lys Ala His Glu Leu GlyIle Val Val 420 425 430 Leu Met Asp Ile Val His Ser His Ala Ser Asn AsnThr Leu Asp Gly 435 440 445 Leu Asn Met Phe Asp Gly Thr Asp Ser Cys TyrPhe His Ser Gly Ala 450 455 460 Arg Gly Tyr His Trp Met Trp Asp Ser ArgLeu Phe Asn Tyr Gly Asn 465 470 475 480 Trp Glu Val Leu Arg Tyr Leu LeuSer Asn Ala Arg Trp Trp Leu Asp 485 490 495 Glu Phe Lys Phe Asp Gly PheArg Phe Asp Gly Val Thr Ser Met Met 500 505 510 Tyr Thr His His Gly LeuSer Val Gly Phe Thr Gly Asn Tyr Glu Glu 515 520 525 Tyr Phe Gly Leu AlaThr Asp Val Asp Ala Val Val Tyr Leu Met Leu 530 535 540 Val Asn Asp LeuIle His Gly Leu Phe Pro Asp Ala Ile Thr Ile Gly 545 550 555 560 Glu AspVal Ser Gly Met Pro Thr Phe Xaa Ile Pro Val Gln Asp Gly 565 570 575 GlyVal Gly Phe Asp Tyr Arg Leu His Met Ala Ile Ala Asp Lys Trp 580 585 590Ile Glu Leu Leu Lys Lys Arg Asp Glu Asp Trp Arg Val Gly Asp Ile 595 600605 Val His Thr Leu Thr Asn Arg Arg Trp Ser Glu Lys Cys Val Ser Tyr 610615 620 Ala Glu Ser His Asp Gln Ala Leu Val Gly Asp Lys Thr Ile Ala Phe625 630 635 640 Trp Leu Met Glu Lys Asp Met Tyr Asp Phe Met Ala Leu AspArg Xaa 645 650 655 Ser Thr Ser Leu Ile Asp Arg Gly Ile Ala Leu His LysMet Ile Arg 660 665 670 Leu Val Thr Met Gly Leu Gly Gly Glu Gly Tyr LeuAsn Phe Met Gly 675 680 685 Asn Glu Phe Gly His Pro Glu Trp Ile Asp PhePro Arg Ala Glu Gln 690 695 700 His Leu Ser Asp Gly Ser Val Ile Pro GlyAsn Gln Phe Ser Tyr Asp 705 710 715 720 Lys Cys Arg Arg Arg Phe Asp LeuGly Asp Ala Glu Tyr Leu Arg Tyr 725 730 735 Arg Gly Leu Gln Glu Phe AspArg Ala Met Gln Tyr Leu Glu Asp Lys 740 745 750 Tyr Glu Phe Met Thr SerGlu His Gln Phe Ile Ser Arg Lys Asp Glu 755 760 765 Gly Asp Arg Met IleVal Phe Glu Lys Gly Asn Leu Val Phe Val Phe 770 775 780 Asn Phe His TrpThr Lys Ser Tyr Ser Asp Tyr Arg Ile Gly Cys Leu 785 790 795 800 Lys ProGly Lys Tyr Lys Val Ala Leu Asp Ser Asp Asp Pro Leu Phe 805 810 815 GlyGly Phe Gly Arg Ile Asp His Asn Ala Glu Tyr Phe Thr Phe Glu 820 825 830Gly Trp Tyr Asp Asp Arg Pro Arg Ser Ile Met Val Tyr Ala Pro Ser 835 840845 Arg Thr Ala Val Val Tyr Ala Leu Val Asp Lys Glu Glu Glu Glu Glu 850855 860 Glu Glu Val Ala Val Val Glu Glu Val Val Val Glu Glu Glu 865 870875 3 1393 DNA Unknown Description of Unknown OrganismbeII gene fragment(branching enzyme II) from Solanum tuberosum (potato) 3 t ctg cca aataat gtg gat ggt tct cct gca att cct cat ggg tcc aga 49 Leu Pro Asn AsnVal Asp Gly Ser Pro Ala Ile Pro His Gly Ser Arg 1 5 10 15 gtg aag atacgt atg gac act cca tca ggt gtt aag gat tcc att cct 97 Val Lys Ile ArgMet Asp Thr Pro Ser Gly Val Lys Asp Ser Ile Pro 20 25 30 gct tgg atc aactac tct tta cag ctt cct gat gaa att cca tat aat 145 Ala Trp Ile Asn TyrSer Leu Gln Leu Pro Asp Glu Ile Pro Tyr Asn 35 40 45 gga ata tat tat gatcca ccc gaa gag gag agg tat atc ttc caa cac 193 Gly Ile Tyr Tyr Asp ProPro Glu Glu Glu Arg Tyr Ile Phe Gln His 50 55 60 cca cgg cca aag aaa ccaaag tcg ctg aga ata tat gaa tct cat att 241 Pro Arg Pro Lys Lys Pro LysSer Leu Arg Ile Tyr Glu Ser His Ile 65 70 75 80 gga atg agt agt ccg gagcct aaa att aac tca tac gtg aat ttt aga 289 Gly Met Ser Ser Pro Glu ProLys Ile Asn Ser Tyr Val Asn Phe Arg 85 90 95 gat gaa gtt ctt cct cgc ataaaa aag ctt ggg tac aat gcg gtg caa 337 Asp Glu Val Leu Pro Arg Ile LysLys Leu Gly Tyr Asn Ala Val Gln 100 105 110 att atg gct att caa gag cattct tat tat gct agt ttt ggt tat cat 385 Ile Met Ala Ile Gln Glu His SerTyr Tyr Ala Ser Phe Gly Tyr His 115 120 125 gtc aca aat ttt ttn gca ccaagc agc cgt ttt gaa acn ccc gac gac 433 Val Thr Asn Phe Xaa Ala Pro SerSer Arg Phe Glu Xaa Pro Asp Asp 130 135 140 ctt aag tct ttg att gat aaagct cat gag cta gga att gtt gtt ctc 481 Leu Lys Ser Leu Ile Asp Lys AlaHis Glu Leu Gly Ile Val Val Leu 145 150 155 160 atg gac att gtt cac agccat gca tca aat aat act tta gat gga ctg 529 Met Asp Ile Val His Ser HisAla Ser Asn Asn Thr Leu Asp Gly Leu 165 170 175 aac atg ttt gac ggc acagat agt tgt tac ttt cac tct gga gct cgt 577 Asn Met Phe Asp Gly Thr AspSer Cys Tyr Phe His Ser Gly Ala Arg 180 185 190 ggt tat cat tgg atg tgggat tcc cgc ctc ttt aac tat gga aac tgg 625 Gly Tyr His Trp Met Trp AspSer Arg Leu Phe Asn Tyr Gly Asn Trp 195 200 205 gag gta ctt agg tat cttctc tca aat gcg aga tgg tgg ttg gat gag 673 Glu Val Leu Arg Tyr Leu LeuSer Asn Ala Arg Trp Trp Leu Asp Glu 210 215 220 ttc aaa ttt gat gga tttaga ttt gat ggt gtg aca tca atg atg tat 721 Phe Lys Phe Asp Gly Phe ArgPhe Asp Gly Val Thr Ser Met Met Tyr 225 230 235 240 act cac cac gga ttatcg gtg gga ttc act ggg aac tac gag gaa tac 769 Thr His His Gly Leu SerVal Gly Phe Thr Gly Asn Tyr Glu Glu Tyr 245 250 255 ttt gga ctc gca actgat gtg gat gct gtt gtg tat ctg atg ctg gtc 817 Phe Gly Leu Ala Thr AspVal Asp Ala Val Val Tyr Leu Met Leu Val 260 265 270 aac gat ctt att catggg ctt ttc cca gat gca att acc att ggt gaa 865 Asn Asp Leu Ile His GlyLeu Phe Pro Asp Ala Ile Thr Ile Gly Glu 275 280 285 gat gtt agc gga atgccg aca ttt tnt att ccc gtt caa gat ggg ggt 913 Asp Val Ser Gly Met ProThr Phe Xaa Ile Pro Val Gln Asp Gly Gly 290 295 300 gtt ggc ttt gac tatcgg ctg cat atg gca att gct gat aaa tgg att 961 Val Gly Phe Asp Tyr ArgLeu His Met Ala Ile Ala Asp Lys Trp Ile 305 310 315 320 gag ttg ctc aagaaa cgg gat gag gat tgg aga gtg ggt gat att gtt 1009 Glu Leu Leu Lys LysArg Asp Glu Asp Trp Arg Val Gly Asp Ile Val 325 330 335 cat aca ctg acaaat aga aga tgg tcg gaa aag tgt gtt tca tac gct 1057 His Thr Leu Thr AsnArg Arg Trp Ser Glu Lys Cys Val Ser Tyr Ala 340 345 350 gaa agt cat gatcaa gct cta gtc ggt gat aaa act ata gca ttc tgg 1105 Glu Ser His Asp GlnAla Leu Val Gly Asp Lys Thr Ile Ala Phe Trp 355 360 365 ctg atg gac aaggat atg tat gat ttt atg gct ctg gat aga ccn tca 1153 Leu Met Asp Lys AspMet Tyr Asp Phe Met Ala Leu Asp Arg Xaa Ser 370 375 380 aca tca tta atagat cgt ggg ata gca ttg cac aag atg att agg ctt 1201 Thr Ser Leu Ile AspArg Gly Ile Ala Leu His Lys Met Ile Arg Leu 385 390 395 400 gta act atggga tta gga gga gaa ggg tac cta aat ttc atg gga aat 1249 Val Thr Met GlyLeu Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn 405 410 415 gaa ttc ggccac cct gag tgg att gat ttc cct agg gct gaa caa cac 1297 Glu Phe Gly HisPro Glu Trp Ile Asp Phe Pro Arg Ala Glu Gln His 420 425 430 ctc tct gatggc tca gta att ccc gga aac caa ttc agt tat gat aaa 1345 Leu Ser Asp GlySer Val Ile Pro Gly Asn Gln Phe Ser Tyr Asp Lys 435 440 445 tgc aga cggaga ttt gac ctg gga gat gca gaa tat tta aga tac cgt 1393 Cys Arg Arg ArgPhe Asp Leu Gly Asp Ala Glu Tyr Leu Arg Tyr Arg 450 455 460 4 464 PRTUnknown Description of Unknown OrganismbeII gene fragment (branchingenzyme II) from Solanum tuberosum (potato) 4 Leu Pro Asn Asn Val Asp GlySer Pro Ala Ile Pro His Gly Ser Arg 1 5 10 15 Val Lys Ile Arg Met AspThr Pro Ser Gly Val Lys Asp Ser Ile Pro 20 25 30 Ala Trp Ile Asn Tyr SerLeu Gln Leu Pro Asp Glu Ile Pro Tyr Asn 35 40 45 Gly Ile Tyr Tyr Asp ProPro Glu Glu Glu Arg Tyr Ile Phe Gln His 50 55 60 Pro Arg Pro Lys Lys ProLys Ser Leu Arg Ile Tyr Glu Ser His Ile 65 70 75 80 Gly Met Ser Ser ProGlu Pro Lys Ile Asn Ser Tyr Val Asn Phe Arg 85 90 95 Asp Glu Val Leu ProArg Ile Lys Lys Leu Gly Tyr Asn Ala Val Gln 100 105 110 Ile Met Ala IleGln Glu His Ser Tyr Tyr Ala Ser Phe Gly Tyr His 115 120 125 Val Thr AsnPhe Xaa Ala Pro Ser Ser Arg Phe Glu Xaa Pro Asp Asp 130 135 140 Leu LysSer Leu Ile Asp Lys Ala His Glu Leu Gly Ile Val Val Leu 145 150 155 160Met Asp Ile Val His Ser His Ala Ser Asn Asn Thr Leu Asp Gly Leu 165 170175 Asn Met Phe Asp Gly Thr Asp Ser Cys Tyr Phe His Ser Gly Ala Arg 180185 190 Gly Tyr His Trp Met Trp Asp Ser Arg Leu Phe Asn Tyr Gly Asn Trp195 200 205 Glu Val Leu Arg Tyr Leu Leu Ser Asn Ala Arg Trp Trp Leu AspGlu 210 215 220 Phe Lys Phe Asp Gly Phe Arg Phe Asp Gly Val Thr Ser MetMet Tyr 225 230 235 240 Thr His His Gly Leu Ser Val Gly Phe Thr Gly AsnTyr Glu Glu Tyr 245 250 255 Phe Gly Leu Ala Thr Asp Val Asp Ala Val ValTyr Leu Met Leu Val 260 265 270 Asn Asp Leu Ile His Gly Leu Phe Pro AspAla Ile Thr Ile Gly Glu 275 280 285 Asp Val Ser Gly Met Pro Thr Phe XaaIle Pro Val Gln Asp Gly Gly 290 295 300 Val Gly Phe Asp Tyr Arg Leu HisMet Ala Ile Ala Asp Lys Trp Ile 305 310 315 320 Glu Leu Leu Lys Lys ArgAsp Glu Asp Trp Arg Val Gly Asp Ile Val 325 330 335 His Thr Leu Thr AsnArg Arg Trp Ser Glu Lys Cys Val Ser Tyr Ala 340 345 350 Glu Ser His AspGln Ala Leu Val Gly Asp Lys Thr Ile Ala Phe Trp 355 360 365 Leu Met AspLys Asp Met Tyr Asp Phe Met Ala Leu Asp Arg Xaa Ser 370 375 380 Thr SerLeu Ile Asp Arg Gly Ile Ala Leu His Lys Met Ile Arg Leu 385 390 395 400Val Thr Met Gly Leu Gly Gly Glu Gly Tyr Leu Asn Phe Met Gly Asn 405 410415 Glu Phe Gly His Pro Glu Trp Ile Asp Phe Pro Arg Ala Glu Gln His 420425 430 Leu Ser Asp Gly Ser Val Ile Pro Gly Asn Gln Phe Ser Tyr Asp Lys435 440 445 Cys Arg Arg Arg Phe Asp Leu Gly Asp Ala Glu Tyr Leu Arg TyrArg 450 455 460

We claim:
 1. A vector which contains an isolated DNA sequence encodingstarch branching enzyme II (SBE II) of potato, wherein the DNA sequenceis in the antisense orientation in relation to a promoter operable in aplant which is operably linked to said DNA sequence, wherein said DNAaffects the amylose/amylopectin ratio in potato starch and the patternof amylopectin branching when expressed in a potato plant.
 2. A processfor the production of transgenic potatoes with a reduced degree ofbranching of amylopectin starch, which comprises the following steps: a)transfer and incorporation of a vector according to claim 1 into thegenome of a potato cell, and b) regeneration of intact, whole plantsfrom the transformed cells.
 3. A process according to claim 2, whereinthe vector also comprises an antisense construct of starch branchingenzyme I (SBE I).
 4. A process according to claim 2, wherein the vectoralso comprises an antisense construct of potato granule bound starchsynthase II.
 5. A process according to claim 3, wherein the vector alsocomprises an antisense construct of potato granule bound starch synthaseII.
 6. A process according to claim 2, wherein the vector also comprisesan antisense construct of potato starch disproportionating enzyme(D-enzyme).
 7. A process according to claim 3, wherein the vector alsocomprises an antisense construct of potato starch disproportionatingenzyme (D-enzyme).
 8. A process according to claim 4, wherein the vectoralso comprises an antisense construct of potato starchdisproportionating enzyme (D-enzyme).
 9. A process according to claim 2,wherein the vector also comprises an antisense construct of potatostarch debranching enzyme.
 10. A process according to claim 3, whereinthe vector also comprises an antisense construct of potato debranchingenzyme.
 11. A process according to claim 4, wherein the vector alsocomprises an antisense construct of potato debranching enzyme.
 12. Aprocess according to claim 6, wherein the vector also comprises anantisense construct of potato debranching enzyme.
 13. A transgenicpotato obtainable by the process according to claim
 2. 14. A transgenicpotato obtainable by the process according to claim
 3. 15. A transgenicpotato obtainable by the process according to claim
 4. 16. A transgenicpotato obtainable by the process according to claim
 6. 17. A transgenicpotato obtainable by the process according to claim
 9. 18. A process forisolating starch or starch proteins from the transgenic potato of claim13, comprising: i) homogenizing said transgenic potato to create ahomogenate; ii) obtaining a juice fraction from said homogenate; iii)allowing the starch to sediment from said juice fraction; and iv)collecting the starch.
 19. A process for isolating starch or starchproteins from the transgenic potato of claim 14, comprising: i)homogenizing said transgenic potato to create a homogenate; ii)obtaining a juice fraction from said homogenate; iii) allowing thestarch to sediment from said juice fraction; and iv) collecting thestarch.
 20. A process for isolating starch or starch proteins from thetransgenic potato of claim 15, comprising: i) homogenizing saidtransgenic potato to create a homogenate; ii) obtaining a juice fractionfrom said homogenate; iii) allowing the starch to sediment from saidjuice fraction; and iv) collecting the starch.
 21. A process forisolating starch or starch proteins from the transgenic potato of claim16, comprising: i) homogenizing said transgenic potato to create ahomogenate; ii) obtaining a juice fraction from said homogenate; iii)allowing the starch to sediment from said juice fraction; and iv)collecting the starch.
 22. A process for isolating starch or starchproteins from the transgenic potato of claim 17, comprising: i)homogenizing said transgenic potato to create a homogenate; ii)obtaining a juice fraction from said homogenate; iii) allowing thestarch to sediment from said juice fraction; and iv) collecting thestarch.
 23. The vector of claim 1, which comprises a DNA having SEQ IDNO:1 in antisense orientation.
 24. The vector of claim 1, wherein saidpromoter is selected from the group consisting of potato granule-boundstarch synthase I, or a promoter derived from the potato starchbranching enzyme I or II gene.